How can i extract DNA from hemolysis blood - (Aug/24/2008 )

Hi all..please help me. i was used Qiagen extraction kit for exrtraction DNA from hemolysed blood but in final step when i add AE buffer to elute DNA from column change the color to yellowish. what is the source of that color and how removed ?

please help me

thank you for all

-aleemy2004-

I don't have experience extracting genomic DNA from blood via columns. However I have a feeling that the colouring comes contaminants from the lysed redblood cells that have become bound to the silica of the column.

Have you confirmed that you have DNA in your yellow coloured sample? Can it do what you want it to do (i assume PCR). It is sometimes difficult to remove all the contaminating compounds (especially if the DNA is extracted from animal tissue). And you have to just live with it.

But if you want and the sample does contain DNA, you could try doubling the wash steps. Do the AW1 wash twice, followed by the AW2 wash which is also repeated twice.

If worse comes to worse, you can used phenol-chloroform extraction (as per the traditional method) and clean up the DNA sample that way.

-perneseblue-

Again, no experience of hemolysed blood but yellow could be breakdown product of hemoglobin.

Pernesblue, I extract from mouse tail clipping for genotyping and use DNA Purification Kit. Can't that be used for purification of DNA from other animal tissues or blood in this example?

-Bungalow Boy-

I believe it should. In theory the silicon column should extract DNA from most tissue. Although some tissue will take more work to lyse the cells.

However given my inexperience with columns used to extract genomic DNA, I am not so sure about blood (unless you use the white buffy layer... unlike heamolysed blood), liver and adipose tissue. Liver and blood have high levels of glycogen/carbohydrate compounds, (adipose tissue, lipids) and they will take some washing to removed from the column. I am not sure if the required wash solutions come with the standard DNA genomic extraction kit.

-perneseblue-

QUOTE (perneseblue @ Aug 25 2008, 12:34 AM)

I believe it should. In theory the silicon column should extract DNA from most tissue. Although some tissue will take more work to lyse the cells.

However given my inexperience with columns used to extract genomic DNA, I am not so sure about blood (unless you use the white buffy layer... unlike heamolysed blood), liver and adipose tissue. Liver and blood have high levels of glycogen/carbohydrate compounds, (adipose tissue, lipids) and they will take some washing to removed from the column. I am not sure if the required wash solutions come with the standard DNA genomic extraction kit.